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Updated: Sep 9, 2025

Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Composition and Structure of Gut Microbiota of Wild and Captive Epinephelus morio via 16S rRNA Analysis and
Grecia Montalvo-Fernández1, Joanna M Ortiz-Alcantara1, Claudia Durruty-Lagunes2
1Laboratorio de Estudios Ecogenómicos, Facultad de Ciencias, Universidad Nacional Autónoma de México (UNAM), Parque Científico Tecnológico de Yucatán, Carretera Sierra Papacal-Chuburná Puerto, Km, 5.5, Tablaje 31257, Sierra Papacal, Mérida 97302, Yucatán, Mexico.
Abstract:
The gut microbiota plays an essential role in the host's metabolism. Its composition and structure depend on biological and environmental factors. This work was designed to identify the composition and structure of the wild and captive red grouper (Epinephelus morio) microbiota and make predictions regarding its metabolic functions. Our hypothesis stated that wild and captive individuals would share the most abundant taxonomic groups, forming a core microbiota, and individuals in captivity might have exclusive taxonomic groups. Metagenomic DNA was extracted from the intestinal contents of wild and captive individuals. The 16S rRNA gene was amplified and sequenced using Illumina pair-end technology. QIIME2 pipeline was used for sequence analysis and alpha and beta diversity assessment. PICRUSt was used to infer metabolic functions. Twenty-nine phyla were identified; the most abundant were Pseudomonadota, Bacillota, Fusobacteriota, and Actinomycetota. The dominant genera were Photobacterium, Vibrio, Cetobacterium, and Escherichia-Shigella. The metabolic prediction analysis suggested that the Epinephelus morio gut microbiota is related to food digestion, the immune system, antioxidant enzymes, antibiotic resistance, and vitamin B12 transport. We concluded that the microbiota of E. morio established in captivity is sensitive to environmental changes such as water pollution, which can cause a decrease in diversity.
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